This invention relates to heterocyclic ring-fused pyrimidine derivatives and methods of using the same in the treatment of hyperproliferative diseases, such as cancers and acnes, in mammals.
Many of the current treatment regimes for cancer utilize compounds which inhibit DNA synthesis. Such compounds are toxic to cells generally but their toxic effect on the rapidly dividing tumor cells can be beneficial. Alterative approaches to anti-cancer agents which act by mechanisms other than the inhibition of DNA synthesis have been explored in order to enhance the selectivity of action against cancer cells.
It is known that a cell may become cancerous by virtue of the transformation of a portion of its DNA into an oncogene (i.e. a gene which, on activation, leads to the formation of malignant tumor cells). Many oncogenes encode proteins which are aberrant tyrosine kinases capable of causing cell transformation. Alternatively, the overexpression of a normal proto-oncogenic tyrosine kinase may also result in proliferative disorders, sometimes resulting in a malignant phenotype.
Receptor tyrosine kinases are large enzymes which span the cell membrane and possess an extracellular binding domain for growth factors such as epidermal growth factor, a transmembrane domain, and an intracellular portion which functions as a kinase to phosphorylate specific tyrosine residues in proteins and hence to influence cell proliferation. It is known that such kinases are frequently aberrantly expressed in common human cancers such as breast cancer, gastrointestinal cancer such as colon, rectal or stomach cancer, leukemia, and ovarian, bronchial or pancreatic cancer. It has also been shown that epidermal growth factor receptor (EGFR) which possesses tyrosine kinase activity is mutated and/or overexpressed in many human cancers such as brain, lung, squamous cell, bladder, gastric, breast, head and neck, oesophageal, gynecological and thyroid tumors.
Accordingly, it has been recognized that inhibitors of receptor tyrosine kinases are useful as a selective inhibitors of the growth of mammalian cancer cells. For example, erbstatin, a tyrosine kinase inhibitor selectively attenuates the growth in athymic nude mice of a transplanted human mammary carcinoma which expresses epidermal growth factor receptor tyrosine kinase (EGFR) but is without effect on the growth of another carcinoma which does not express the EGF receptor.
Various other compounds, such as styrene derivatives, have also been shown to possess tyrosine kinase inhibitory properties. More recently three European patent publications, namely EP 0 566 226 A1, EP 0 602 851 A1 and EP 0 520 722 A1 have disclosed that certain heteroaryl-fused pyrimidine derivatives possess anti-cancer properties which result from their tyrosine kinase inhibitory properties. Also PCT publication WO 92/20642 discloses bis-mono and bicyclic aryl and heteroaryl compounds as tyrosine kinase inhibitors.
European patent publication EP 0 496 617 A1 discloses certain pyrazolo[3,4-d]pyrimidines and pyrrolo[2,3-d]pyrimidines which possess adenosine kinase inhibitory properties.
European patent publication EP 0 475 413 A2 discloses certain carbocyclic nucleoside analogs as useful immunosuppressants.
European patent publication EP 0 414 386 A1 discloses certain pyrido[2,3-d]pyrimidines as fungicides, insecticides and miticides. The synthesis and antiallergic activity of 9-aryl-8-azaadenine derivatives is described in II Farmcoxe2x80x94Ed. Sc., vol 35, fasc. 4 p308-323 (1980).
Co-pending U.S. patent applications (U.S. Ser. Nos. 08/200,359 and 08/413,800) and PCT application docket no. PC8836A, assigned to the Assignee of this application, describe optionally substituted indolyl- and phenylamino quinazolines, respectively, which are useful in the treatment of hyperproliferative diseases involving receptor tyrosine kinases. In addition U.S. Pat. No. 4,012,513 discloses certain 1-(heterocyclic)-indol-3-yl-acetic acid derivatives that have anti-inflammatory, analgesic and antipyretic activity.
Although the anti-cancer compounds described above make a significant contribution to the art there is a continuing search in this field of art for improved anti-cancer pharmaceuticals.
This invention is directed to compounds of the Formula 
and stereoisomers, pharmaceutically acceptable salts and prodrugs thereof, wherein Y together with the carbons to which it is attached form a 5 or 6 membered, optionally unsaturated or aromatic ring wherein said ring is optionally substituted with (R3)p and/or R4 groups and comprises one to three heteroatoms selected from S, O and N with the proviso that at least one of said hetero atoms is N;
Z is NR1R2 wherein R1 is H and R2 is phenyl substituted by (R5)m or Q or R1R2N is a group of the formula 
wherein the dotted line represents an optional double bond;
each R3is attached to a carbon atom in Y and is independently selected from
a. hydrogen, trifluoromethyl, halo, nitro, hydroxy, amino, cyano, (C1-C4)alkyl, (C1-C4)alkoxy, (C1-C4)alkoxycarbonyl, (C1 -C4)alkanoyloxy, (C1-C4)alkanoylamino, carboxy, phenoxy, benzoyloxy, carbamoyl, mono-N- or di-N-N-di-(C1-C4)alkylcarbamoyl, mono-N- or di-N,N-(C1-C4)alkylamino, mono-N or di-N,N-(hydroxy(C2-C4)alkyl)amino, mono-N or di-N,N-((C1-C4)alkoxy(C2-C4)alkyl)amino, anilino, pyrrolidin-1-yl, piperidin-1-yl, morpholino, piperazin-1-yl, 4(C1-C4)alkylpiperazin-1-yl, pyridyl, pyrrolo, imidazolo, thiazolo, benzimidazolo, pyridonyl, (C1-4)alkylthio, phenylthio, or such groups substituted on (C1-C4)alkyl;
b. hydroxy(C2-C4)alkoxy(C1-C4)alkyl, (C1-C4)alkoxy-(C2-C4)alkoxy-(C1-C4)alkyl, hydroxy(C2-C4)alkylthio(C1-C4)alkyl, (C1-C4)alkoxy(C2-C4)alkylthio(C1-C4)alkyl, hydroxyamino, benzoylamino, mono-N or di- N,N-(C1-C4)alkylcarbamoylmethylamino, carbamoylmethylamino, (C1-C4)alkoxycarbonylamino, (C1-C4)alkanoylamino, carboxymethylamino, (C1-C4)alkoxycarbonylmethylamino, (C1-C4)alkoxyamino, (C2-C4)alkanoyloxyamino, phenyl(C1-C4)alkylamino, (C1-C4)alkylsulphonylamino, benzenesulphonamido, 3phenylureido, 2-oxopyrrolidin-1-yl, 2,5-dioxopyrrolidin-1-yl, ureido,(C1-C4)alkoxy(C1-C4)alkylcarbonylamino,(C1-C4)alkylsulfinyl,(C1-C4)alkylsulfonyl, (C1-C4)alkoxy(C2-C4)alkylthio, mono-, di- or trifluoromethyloxy, (C1-C4)alkylenedioxy, benzyloxy, guanidino, aminocarbonyl, mono-N- or di-N,N-(C1-C4)alkylaminocarbonyl, phenyl(C1-C4)alkoxy, carboxymethoxy, (C1-C4)alkoxycarbonylmethoxy, carbamoylmethoxy, mono-N or di-N,N-(C1-C4)alkyl carbamoylmethoxy, mono-N- or di-N,N-(hydroxy(C2-C4)alkyl)carboxamido, mono-N- or di-N,N-((C1-C4)alkoxy (C2-C4)alkyl)carboxamido or bis((C1-C4)alkanesulfonyl)amido; or
c. (C2-C4)alkoxy, (C2-C4)alkylthio, (C2-C4)alkanoyloxy, (C2-C4)alkylamino, (C1-C4)alkyl(C1-C4)alkylenedioxy, (C2-C4)alkanoylamino, (C2-C4)alkenyl, or (C2-C4)alkynyl; each such group substituted with amino, halo, hydroxy, (C2-C4)alkanoyloxy, (C1-C4)alkoxy, mono-N- or di-N,N-(C1-C4)alkylamino, mono-N or di-N,N-(hydroxy(C2-C4)alkyl)amino, mono-N or di-N,N-((C1-C4)alkoxy(C2-C4)alkyl)amino, (C1-C4)alkanoylamino, phenoxy, anilino, imidazol-1-yl, phenylthio, piperidino, pyridyl, carboxy(C1-C4)alkylthio(C1-C4)alkoxy, morpholino, piperazin-1-yl-, 4-(C1-C4)alkylpiperazin-1-yl-, carboxy, (C1-C4)alkoxycarbonyl, carbamoyl, mono-N- or di-N,N-(C1-C4)alkylcarbamoyl, carboxamido, mono-N- or di-N,N-(C1-C4)alkylcarboxamido or mono-N- or di-N,N-(hydroxy(C2-C4)alkyl)carboxamido; and any phenyl in an R3 substituent is optionally mono- or di- substituted with halo, nitro, trifluoromethyl, hydroxy, (C1-C4)alkoxy, (C1 -C4)alkyl, amino, mono-N-alkylamino, or N,N-dialkylamino;
R4 is attached to a N-atom in Y and is independently selected from:
hydrogen, (C1-C4)alkyl, (C1-C4)alkoxycarbonyl, (C1-C4)alkanoyl, (C1-C4)alkylsulfonyl, arylsulfonyl, allyl; or a (C2-C4)alkyl, (C2-C4)alkanoyl, or (C2-C4)alkoxycarbonyl, (C2-C4)alkylsulfonyl, each such group substituted with amino, halo, hydroxy, (C2-C4)alkanoyloxy, (C1-C4)alkoxy, mono-N- or di-N,N-(C1-C4)alkylamino, mono-N or di-N,N-(hydroxy(C2-C4)alkyl)amino, mono-N or di-N,N-(C1-C4)alkoxy(C2-C4)alkyl)amino, (C1-C4)alkanoylamino, phenoxy, anilino, imidazol-1-yl, phenylthio, piperidino, morpholino, piperazin-1-yl-, 4-(C1-C4)alkylpiperazin-1-yl-, phenyl, pyridyl, pyrrolo, imidazolo, thiazolo, benzimidazolo, pyridenyl, carboxy, (C1-C4)alkoxycarbonyl, carbamoyl, mono-N- or di-N,N-(C1-C4)alkylcarbamoyl, carboxamido, mono-N- or di-N,N-(C1-C4)alkylcarboxamido or mono-N- or di-N,N-(hydroxy(C2-C4)alkyl)carboxamido; and any phenyl in an R4 substituent is optionally mono- or di- substituted with halo, nitro, trifluoromethyl, hydroxy, (C1-C4)alkoxy, (C1-C4)alkyl, amino, mono-N-alkylamino, or N,N-dialkylamino; but specifically R4 is not furanosyl, pyranosyl, or cyclopentyl;
each R5 is independently selected from mono-, di- or tri-fluoromethyl, halo, nitro, hydroxy, amino, azido, isothiocyano, (C1-C4)alkyl, phenyl, thienyl, (C1-C4)alkoxy, benzyloxy, phenoxy, (C2-C6)alkenyl, (C2-C6)alkynyl (C1-C4)alkylenedioxy, cyano, benzoylamino, trifluoromethylcarbonylamino, (C1-C4)alkanoylamino, (C1-C4)alkanoyl, N-mono- or N,N-di-(C1-C4)alkylamino, (C1-C4)alkylsulfonylamino, trifluoromethylsulfonylamino, (C1-C4)alkylthio, (C1-C4)alkylfinyl or (C1-C4)alkylsulfonyl, pyrrol-1-yl, piperidin-1-yl or pyrrolidin-1-yl, said phenyl, benzyloxy, phenoxy and benzoylamino optionally mono-substituted with halo, nitro, trifluoromethyl, hydroxy or (C1-C4)alkyl and said (C1-C4)alkylenedioxy is linked at both ends to adjacent carbons on the benzene moiety;
each R6 is independently selected from hydroxy, amino, N-mono- or N,N-di-(C1-C4)alkylamino, sulfo, or (C1-C4)alkoxy (provided that such groups are not attached to a ring carbon which is directly adjacent to the ring N-), or R6 for each occurrence is independently carboxy, hydroxy(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, amino(C1-C4)alkyl, mono-N- or di-N,N-(C1-C4)alkylamino(C1-C4)alkyl, morpholino (C1-C4)alkyl, 4-(C1-C4)alkyl-piperazin-1-yl(C1-C4)alkyl, carboxy(C1-C4)alkyl, (C1-C4)alkoxycarbonyl, sulfo(C1-C4)alkyl, pyridyl(C1-C4)alkyl or (C1-C4)alkyl;
m is an integer from 1 to 3;
n is 0, 1 or 2;
p is 0 or an integer from 1-3;
with the proviso that when Y, In the direction shown by the arrow in formula I, is xe2x80x94CR3xe2x95x90Nxe2x80x94CR3xe2x95x90CR3xe2x80x94, p=0, m=1 and Z is substituted phenyl then R5 is not 4-ethoxy, 4-methoxy, 4-trifluoromethoxy, 4-t-butyl or 4isopropyl;
Q is a 9- or 10-membered bicyclic heteroaryl cyclic moiety, or a hydrogenated derivative thereof, containing one or two nitrogen heteroatoms and optionally containing a further heteroatom selected from nitrogen, oxygen and sulphur, or Q is a 9- or 10-membered bicyclic aryl moiety, or a hydrogenated derivative thereof, which heterocyclic or aryl moiety, or hydrogenated derivatives thereof, may optionally bear one or two substituents selected from halogeno, hydroxy, oxo, amino, nitro, carbamoyl, (C1-C4)alkyl, (C1-C4)alkoxy, (C1-C4)alkylamino, di-[(C1-C4)alkyl]amino, and (C2-C4)alkanoylamino; with the proviso that when Y, in the direction shown by the arrow in formula I, is xe2x80x94NR4xe2x80x94CR3xe2x95x90CR3xe2x80x94, R3 xe2x95x90CH3 and R4xe2x95x90H, then R5 is not 4-CH3, 3,5-(CH3)2, 2,6-(CH3)2, 2-C2H5, 4C2H5, 4n-C4H9, 2-Cl, 4-Cl, 3,4-Cl2, 2-F, or 3-CF3.
According to another aspect of the invention there is provided a compound as described above wherein Y, in the direction shown by the arrow in formula I is selected from xe2x80x94Nxe2x95x90CR3xe2x80x94NR4xe2x80x94, xe2x80x94CR3xe2x95x90CR3xe2x80x94NR4xe2x80x94, xe2x80x94NRxe2x80x94CR3=CR3xe2x80x94, xe2x80x94Nxe2x95x90Nxe2x80x94NR4xe2x80x94, xe2x80x94NR4xe2x80x94Nxe2x95x90Nxe2x80x94NR4xe2x80x94, NR4xe2x80x94Nxe2x95x90CR3, xe2x95x90CR3xe2x80x94NR4xe2x80x94CR3xe2x95x90, xe2x80x94Nxe2x95x90CR3xe2x80x94CR3xe2x95x90CR3xe2x80x94, xe2x80x94CR3xe2x95x90Nxe2x80x94CR3xe2x95x90CR3xe2x80x94, xe2x80x94CR3xe2x95x90CR3xe2x80x94Nxe2x95x90CR3xe2x80x94, xe2x80x94CR3xe2x95x90CR3xe2x80x94CR3xe2x95x90Nxe2x80x94.
Another aspect of the invention provides a compound as described above wherein each R3 is independently selected from hydrogen, hydroxy, (C1-C4)alkoxy, hydroxy(C2-C4)alkoxy, amino(C2-C4)alkyl, amino(C2-C4)alkoxy, (C1-C4)alkoxy(C2-C4)alkoxy, hydroxy(C1-C4)alkyl(C1-C4)alkylenedioxy, (C1-C4)alkoxy(C1-C4)alkyl(C1-C4)alkylenedioxy, mono-N- or di-N,N-(C1-C4)alkylamino(C2-C4)alkoxy, 3- or 4-(C1-C4)alkoxy-(2-hydroxy)-(C3-C4)alkoxy, carboxy(C1-C4)alkoxy, morpholino(C2-C4)alkoxy, imidazol-1-yl(C2-C4)alkoxy, 4(C1-C4)alkylpiperazin-1-yl-(C2-C4)alkoxy, (C1-C4)alkoxy(C1-C4)alkanoyloxy, nitro, hydroxylamino, amino, phenyl, pyridyl, pyrrolo, imidazolo, thiazolo, benzimidazolo, pyridonyl, mono-N- or di-N,N-(C1-C4)alkylamino, (C1-C4)alkanoylamino, hydroxy(C2-C4)alkylamino, (C1-C4)alkoxy(C2-C4)alkylamino, (C1-C4)alkylsulfonamido, morpholino, (C1-C4)alkyl-piperazin-1-yl, bis(C1-4)alkanesulfonamido, di-N,N-(C1-C4)alkylamino(C2-C4)alkylamino, (C1-C4)alkylamino(C2-C4)alkylamino, piperidin-1-yl, imidazol-1-yl, pyrrolidin-1-yl, (C1-C4)alkoxy(C1-C4)alkylcarbonylamino, carboxy, (C1-C4)alkoxycarbonyl, (C1-C4)alkoxycarbonyl(C1-C4)alkoxy, amido, mono-N- or di-N,N-(C1-C4)alkylaminocarbonyl, mono-N- or di-N,N-(hydroxy(C2-C4)alkyl)aminocarbonyl, (C1-C4)alkyl1, hydroxy(C1-C4)alkyl, mono-N- or di-N,N-((C1-C4)alkoxy(C1-C4)alkyl)amino(C1-C4)alkyl, mono-N- or di-N,N-(C1-C4)alkylamino(C1-C4)alkyl, (C1-C4)alkanoylamino(C1-C4)alkyl, (C1-C4)alkoxy(C2-C4)alkoxy(C1-C4)alkyl, (C1-C4)alkylthio, (C1-C4)alkoxy(C2-C4)alkylthio or hydroxy(C2-C4)alkylthio; and R4 is selected from hydrogen, benzyl, phenyl, a (C2-C4)alkyl, hydroxy(C2-C4)alkyl, or hydroxy(C2-C4)alkyl, amino(C2-C6)alkyl, (C2-C4)alkoxycarbonyl each such group substituted with amino, halo, hydroxy, (C2-C4)alkanoyloxy, (C1-C4)alkoxy, mono-N- or di-N,N-(C1-C4)alkylamino, mono-N or di-N,N-(hydroxy(C2-C4)alkyl)amino, mono-N or di-N,N-((C1-C4)alkoxy(C2-C4)alkyl)amino, sulfonylaryl(C1-C4)alkylamine, (C1-C4)alkanoylamino, imidazol-1-yl, piperidino, morpholino, piperazin-1-yl-, 4(C1-C4)alkylpiperazin-1-yl-, pyridyl, pyrrolo, imidazolo, thiazolo, pyridenyl, carboxy, (C1-C4)alkoxycarbonyl, carbamoyl, mono-N- or di-N,N-(C1-C4)alkylcarbamoyl, carboxamido, mono-N- or di-N,N-(C1-C4)alkylcarboxamido or mono-N- or di-N,N-(hydroxy(C2-C4)alkyl)carboxamido.
Yet another aspect of the invention provides a compound as described above wherein Y in the direction shown by the arrow in formula I is selected from xe2x80x94CR3xe2x95x90CR3xe2x80x94NR4xe2x80x94, xe2x80x94NR4xe2x80x94CR3xe2x95x90CR3xe2x80x94 and xe2x80x94CHxe2x95x90CR3xe2x80x94Nxe2x95x90CHxe2x80x94.
According to another aspect of the invention there is provided a compound as described above wherein Y, in the direction shown by the arrow in formula I is selected from xe2x80x94NR4xe2x80x94CR3xe2x95x90CR3xe2x80x94, or xe2x80x94CHxe2x95x90CR3xe2x80x94Nxe2x95x90CHxe2x80x94 and xe2x80x94CR3xe2x95x90CR3xe2x80x94NR4 xe2x80x94.
Yet another aspect of the invention provides a compound as described above wherein Y, in the direction shown by the arrow in formula 1, is xe2x80x94CR3xe2x95x90CR3xe2x80x94NR4xe2x80x94and R4 is hydrogen.
Another aspect of the invention provides a compound as described above wherein R1R2N is 
and R5, R6, m and n are as defined above.
According to another aspect of the invention there is provided a compound as described above wherein each R5 is Independently selected from 4-hydroxy, 4amino, fluoro, 5-hydroxy, 5-amino, 6-halo, 6methyl, 6-ethenyl, 6-ethynyl, 6-nitro and 7-methyl and each R6 is independently selected from hydroxy, amino, N-mono- or N,N-di-(C1-C4)alkylamino, sulfo, or (C1-C4)alkoxy (provided that such groups are not attached to a ring carbon which is directly adjacent to the ring N-), or R6 for each occurrence is independently carboxy, hydroxy(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, amino(C1-C4)alkyl, mono-N- or di-N,N-(C1-C4)alkylamino(C1-C4)alkyl, morpholino (C1-C4)alkyl, 4-(C1-C4)alkyl-piperazin-1-yl(C1-C4)alkyl, carboxy(C1-C4)alkyl, (C1-C4)alkoxycarbonyl, sulfo(C1-C4)alkyl, pyridyl(C1-C4)alkyl and (C1-C4)alkyl.
Another aspect of the invention provides a compound as described above wherein R1 is H and R2 is (R5)m-substituted phenyl wherein R5 and m are as defined above.
Yet another aspect of the invention provides a compound as described above wherein each R5 is independently selected from 4fluoro-3-chloro, 3-trifluoromethyl, 4-fluoro-3-trifluoromethyl, 3-nitro-4-chloro, 3-nitro-4-fluoro, 4-fluoro-3-bromo, 3-iodo-5-amino, 3-methyl-4-fluoro, 4-amino, 3-fluoro, 3-hydroxy, 3-amino, 3-halo, 3-methyl, 3-ethenyl, 3-ethynyl, 3-nitro and 4-methyl.
According to another aspect of the invention there is provided a compound as described above wherein R1 is H and R2 is Q.
Another aspect of the invention provides a compound as described above wherein Q is selected from pyrrolo 1,2,3,5-tetrahydro-pyrrolo[2,3-f]indole, 4-, 5-, 6-indolyl, 1H-benzimidazol4yl, 1H-benzimidazol-5-yl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, 1H-indazol-7-yl, 1H-benzotrizol-4-yl, 1H-benzotrizol-5-yl, 1H-benzotrizol-6-yl, 5- or 6-benzoxazolyl, 5- or 6-benzothiazolyl, benzo[c][2,1,3]thiadiazol-4-yl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 4-, 5-, 6-, 7- or 8cinnolinyl, 5-, 6-, 7- or 8-quinazolinyl, or 2-, 5-, or 6-quinoxalinyl, which may optionally bear one or two substituents selected from fluoro, bromo, chloro, methyl, ethyl, ethenyl, ethynyl and methoxy.
Yet another aspect of the invention provides a compound as described above wherein Q is selected from pyrrolo 5-indolyl, 1H-indazol-5-yl, 1H-benzotriazol-5-yl, 6-benzothiazolyl, benzo[c][2,1,3]thiadiazol-4-yl, 5quinolyl, 6quinolyl, 8quinolyl, 5-isoquinolyl, or 5quinocalinyl, which may optionally bear one or two substituents selected from fluoro, bromo, chloro, methyl, ethyl, ethenyl, ethynyl and methoxy.
Preferred compounds of formula I are selected from the group consisting of
(3-ethynyl-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine hydrochloride;
(3-chloro-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine hydrochloride;
4-(6-chloro-2,3-dihydro-indol-1-yl)7H-pyrrolo[2,3-d]pyrimidine hydrochloride;
(7H-pyrrolo[2,3-d]pyrimidin-4-yl)m-tolyl-amine hydrochloride;
(1H-indol-5-yl)-(7H-pyrrolo [2,3-d]pyrimidin-4-yl)-amine hydrochloride;
(6-methylindolin-1-yl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(benzo[b]thien-5-yl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(6-chloro-5-fluoroindolin-1-yl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(1H-indazol-5-yl)-(7H-pyrrolo[2,3]pyrimidin-4-yl)-amine;
1-(4-m-tolylamino-pyrrolo[2,3-d]pyrimidin-7-yl)-ethanone hydrochloride;
(5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-m-tolyl-amine;
(3-chloro-phenyl)-(1H-[1,2,3]triazolo[4,5-d]pyrimidin-7-yl)-amine hydrochloride:
(3-chloro-phenyl-pyrido[4,3-d]pyrimidin-4-yl-amine hydrochloride;
(1H-indol-5-yl)-pyrido[4,3-d]pyrimidin-4-yl-amine hydrochloride;
(3-ethynylphenyl)-(7-methyl-pyrido[4,3-d]pyrimidin-4-yl)-amine hydrochloride;
(3-chloro-phenyl)-(7-methyl-pyrido[4,3-d]pyrimidin-4-yl)-amine hydrochloride;
(3-ethynyl-phenyl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine hydrochloride;
(6-bromo-5-fluoroindolin-1-yl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine;
(6-chloro-5-fluoroindolin-1-yl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine;
(1H-indazol-5-yl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine;
(3-methyl-4-hydroxyphenyl)(6methylpyrido[4,3-d]pyrimidin-4-yl)-amine;
(6-iodoindolin-1-yl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine;
(benzo[b]thien-5-yl)-(pyrido[4,3-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-(9H-purin-6-yl)-amine;
(1H-indol-5-yl)-(9H-purin-6-yl)-amine hydrochloride;
(3-chloro-phenyl)-(9H-purin-6-yl)-amine hydrochloride;
4-(6-chloro-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;
(pyrido[3,4-d]pyrimidin-4-yl)-(m-tolyl)-amine;
(1H-indazol-5-yl)-(pyrido[3,4-d]pyrimidin-4-yl)-amine;
(1H-indol-5-yl)-(pyrido[3,4-d]pyrimidin-4-yl)-amine;
(phenyl)-(pyrido[2,3-d]pyrimidin-4-yl)-amine;
(3-chloro-phenyl)-(pyrido[2,3-d]pyrimidin-4-yl)-amine;
(3-chloro-phenyl)-(pyrido[3,4-d]pyrimidin-4-yl)-amine;
(3-bromo-phenyl)-(pyrido[3,4-d]pyrimidin-4-yl)-amine;
(phenyl)-(pyrido[3,4-d]pyrimidin-4-yl)-amine;
4-(6-chloro-2,3dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;
(pyrido[3,4-d]pyrimidin-4-yl)-(m-tolyl)-amine;
(1H4-indazol-5-yl)-pyrido[3,4-d]pyrimidin-4-yl-amine;
(1H-indol-5-yl)-pyrido[3,4-d]pyrimidin-4-yl-amine;
phenyl-pyrido[2,3-d]pyrimidin-4-yl-amine;
(3-chloro-phenyl-pyrido[2,3-d]pyrimidin-4-yl-amine;
(3-chloro-phenyl-pyrido[3,4-d]pyrimidin-4-yl-amine;
(3-bromo-phenyl-pyrido[3,4-d]pyrimidin-4-yl-amine;
phenyl-pyrido[3,4-d]pyrimidin-4-yl-amine;
(7-benzenesulfonyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-(3ethynyl-phenyl)-amine;
4-(6-chloro-2,3-dihydro-indol-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-6-ol;
(3-ethynyl-phenyl)-[7-(2-morpholin-4-yl-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine;
(3-ethynyl-phenyl)-[7-(2-methoxy-ethyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-amine;
(3-ethynyl-phenyl)-{7-[2-(2-mothoxy-ethoxy)-ethyl]-7H-pyrrolo[2,3-d]pyrimidin-4-yl}-amine;
(7-allyl-pyrrolo[2,3-d]pyrimidin-4-yl)-(3-ethynyl-phenyl)-amine hydrochloride;
(3-ethynyl-phenyl)-(7-methyl-pyrrolo[2,3-d]pyrimidin-4yl)-amine hydrochloride;
(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-(3-ethynyl-phenyl)-amine;
(3-ethynyl-phenyl)-(6-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
4-(3-ethynyl-phenylamino)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid;
(3-ethynyl-phenyl)-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine hydrochloride;
N-(5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N-m-tolyl-acetamide;
4-(3-ethynyl-phenylamino)-7H-pyrrolo[2,3-d]pyrimidin-5-carboxylic acid methyl esterhydrochloride;(3-ethynyl-phenyl)-(5-cyano-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(1H-indazol-5-yl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride; benzo[b]thiophen-5-yl-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride;
(3-ethynyl-4-fluoro-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
2-methyl-4-(6-methyl-pyrido[3,4-d]pyrimidin-4-ylamino)-phenol dihydrochloride;
4-(4-bromo-7-methyl-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine hydrochloride;
4-(6-bromo-7-methyl-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine hydrochloride;
4-(6-bromo-6-fluoro-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine hydrochloride;
(3-chloro-4-fluoro-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride;
(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-(3-trifluoromethyl-phenyl)-amine hydrochloride;
(4-fluoro-3-methyl-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride;
2-iodo-4-(6-methyl-pyrido[3,4-d]pyrimidin-4-ylamino)-phenol hydrochloride;
(4-bromo-3-fluoro-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride;
4-(6,7-dimethyl-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine hydrochloride;
(3ethynyl-phenyl)-pyrido[3,4-d]pyrimidin-4-yl-amine hydrochloride;
benzo[b]thiophen-5-yl-pyrido[3,4-d]pyrimidin-4-yl-amine hydrochloride;
(3-ethynyl-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine hydrochloride; 4-(6-chloro-2,3-dihydro-indol-1-yl)-6-methyl-pyrido[3,4-d]pyrimidine;
(3-ethynyl-phenyl)-(5-methylsulfanyl-7H-pyrrolo[2,3]pyrimidin-4-yl)-amine; and
(1H-indol-5-yl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine methanesulfonate.
Most preferred compounds of the formula I as described above are selected from
(1H-indol-5-yl)-(methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(3-chloro-phenyl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino;
4-(3-ethynyl-phenylamino)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid methyl ester;
4-(3-ethynyl-phenylamino)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile;
(1H-indol-5-yl)-pyrido[3,4-d]pyrimidin-4-yl-amine;
(3chloro-4-fluoro-phenyl)-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
benzo[b]thiophen-5-yl-(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-pyrido[3,4-d]pyrimidin-4-yl-amine;
(4-fluoro-3-methyl-phenyl)-(6methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
4-(6chloro-2,3-dihydro-indol-1-yl)-pyrido[3,4-d]pyrimidine;
pyrido[3,4-d]pyrimidin-4-yl-m-tolyl-amine;
(6-methyl-pyrido[3,4-d]pyrimidin-4-yl)-(3-trifluoromethyl-phenyl)-amine;
(1H-indazol-5-yl)-(6methyl-pyrido[3,4-d]pyrimidin-4-yl)-amine;
(3-ethynyl-phenyl)-(5-methylsulfonyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;
(1H-indol-5-yl)-(7H-pyrrolo[2,3-d]pyrimidin 4-yl)-amine;
(5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-(3-ethynyl-phenyl)-amine;
The invention also provides a compound of the formula 
wherein W, in the direction indicated by the arrows, is selected from xe2x80x94CH2xe2x95x90C(CH3)xe2x80x94Nxe2x95x90CH2xe2x80x94, xe2x80x94CH2xe2x95x90Nxe2x80x94C(CH3)xe2x95x90CH2xe2x80x94 and xe2x95x90C(CH3)xe2x80x94NHxe2x80x94CHxe2x95x90.
According to another aspect of the invention there is provided a compound as described above wherein W is xe2x80x94CH2xe2x95x90C(CH3)xe2x80x94Nxe2x95x90CH2xe2x80x94.
Another aspect of the invention provides a compound as described above wherein W is xe2x80x94CH2xe2x95x90Nxe2x80x94C(CH3)xe2x95x90CH2xe2x80x94.
Yet another aspect of the invention provides a compound as described above wherein W is xe2x95x90C(CH3)xe2x80x94NHxe2x80x94CHxe2x95x90.
The invention also provides a method of treating hyperproliferative disorders which comprises administering to a mammal in need of such treatment a hyperproliferative disorder treating amount of a compound of formula I.
According to another aspect of the invention there is provided a method as described above wherein the hyperproliferative disease is cancer.
Yet another aspect of the invention provides a method as described above wherein the disease is brain, lung, squamous cell, bladder, gastric, pancreatic, hepatic, renal, colorectal, breast, head, neck, oesophageal, gynecological or thyroid cancer.
Another aspect of the Invention provides a method as described above wherein the hyperproliferative disorder is noncancerous.
Another aspect of the invention provides a compound as described above wherein the noncancerous hyperproliferative disorder is psoriasis or benign prostatic hyperplasia.
The invention further provides a pharmaceutical composition for the treatment of hyperproliferative disorder in a mammal which comprises a hyperproliferative disease treating amount of a compound of formula I and a pharmaceutically acceptable carrier.
In the present application certain terms are defined as follows:
By halo is meant chloro, bromo, iodo, or fluoro.
By alkyl is meant straight chain or, when C3 or larger, a cyclic or, when C2 or larger, a branched saturated hydrocarbon.
As used herein, the expression xe2x80x9creaction-inert solventsxe2x80x9d refers to a solvent which does not interact with starting materials, reagents, intermediates or products in a manner which adversely affects the yield of the desired product.
Other features and advantages will be apparent from the specification and claims which describe the invention.
The Formula I compounds, or pharmaceutically acceptable salts and prodrugs thereof can be prepared by any process known to be applicable to the preparation of chemically-related compounds. 
As shown in the Scheme the Formula I compounds can, generally, be made prepared from the 4-chloro or hydroxy derivatives of the appropriately substituted heteroaryl-fused pyrimidine 1 using the appropriately substituted amine ZH 2.
Typically the appropriately substituted 4-haloheteroaryl-fused pyrimidine 1 (or a heteroaryl-fused pyrimidine bearing a suitable displaceable leaving group in the 4-position such as aryloxy, alkyl sulfinyloxy such as trifluoromethanesulfonyloxy, arylsulfinyloxy, siloxy, cyano, pyrazolo, triazolo ortetrazolo), preferably a haloheteroaryl such as 4-chloroheteroaryl derivative, is reacted with the appropriate amine 2 in a solvent such as a (C1-C6)alcohol, dimethylformamide (DMF), N-methylpyrrolidin-2-one, chloroform, acetonitrile, tetrahydrofuran (THF), dimethylsulfoxide (DMSO), 1-4 dioxane, pyridine or other aprotic solvent. The combination can be effected in the presence of a base, preferably an alkali or alkaline earth metal carbonate or hydroxide or a tertiary amine base, such as pyridine, 2,6-lutidine, collidine, N-methyl-morpholine, triethylamine, diethyl isopropyl amine, 4-dimethylamino-pyridine or N,N-dimethylaniline. These bases are hereinafter referred to as xe2x80x9csuitable basesxe2x80x9d. The mixture is maintained at a temperature of from ambient to reflux, preferably from about 35xc2x0 C. to reflux, until substantially no remaining 4-haloheteroaryl-fused pyrimidine can be detected, typically about 2 hours to about 72 hours. The reaction is preferably performed under an inert atmosphere such as dry nitrogen gas.
Generally, the reactants are combined stoichiometrically when a suitable amine base is used, although, for those compounds where a salt (typically the HCl salt) of the amine is used, it is preferable to use excess amine 2, generally an extra equivalent of the amine 2. Alternatively, if an amine base is not used an excess of the amine reactant may be used)
For those compounds where a sterically hindered amine (such as a 2-alkylindoline) or very reactive 4-haloheteroaryl-fused pyrimidine is used it is preferable to use t-butyl alcohol or a polar aprotic solvent such as dimethylformamide or N-methylpyrrolidin-2-one as the solvent.
Other Formula I compounds may be prepared by the following appropriate reactions subsequent to the above coupling.
Compounds of Formula I wherein R3 or R5 is a primary amino or hydroxyamino may be prepared by the reduction of Formula I compounds wherein R3 or R5 is a nitro group.
The reduction may conveniently be carried out by any of the many procedures known for such transformations, The reduction may be carried out, for example, by the hydrogenation of a solution of the nitro compound in a reaction-inert solvent in the presence of a suitable metal catalyst such as palladium or platinum. A further suitable reducing agent is, for example, an activated metal such as activated iron (produced by washing iron powder with a dilute solution of an acid such as hydrochloric acid). Thus, for example, the reduction may be carried out by heating a mixture of the nitro compound and the activated metal in a solvent such as a mixture of water and an alcohol, for example, methanol or ethanol, to a temperature in the range, for example, 50 to 150xc2x0 C., conveniently at or near 70xc2x0 C.
For the production of those compounds of Formula I wherein R5 or R6 incorporates a primary or secondary amino moiety (other than the amino group intended to react with the quinazoline), such free amine is preferably protected prior to the above described reaction followed by deprotection, subsequent to the above described reaction with 4-haloquinazoline.
For a description of protecting groups and their use, see T. W. Greene and P. G. M. Wuts, xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d Second Ed., John Wiley and Sons, New York, 1991.
Nitrogen protecting groups are well known in the art, including (C1-C6)alkoxycarbonyl, optionally substituted benzyloxycarbonyl, aryloxycarbonyl, tributyl, vinyloxycarbonyl, O-nitrophenylsulfonyl, diphenylphosphinyl, p-toluenesulfonyl, and benzyl. The addition of the nitrogen protecting group may be carried out in a chlorinated hydrocarbon solvent such as methylene chloride or 1,2-dichloroethane, or an ethereal solvent such as glyme, diglyme or THF, in the presence or absence of a tertiary amine base such as triethylamine, diisopropylethylamine or pyridine, preferably triethylamine, at a temperature from about 0xc2x0 C. to about 50xc2x0 C., preferably about ambient temperature. Alternatively, the protecting groups are conveniently attached using Schotten-Baumann conditions.
Subsequent to the above described amine coupling reaction the protecting group may be removed by deprotecting methods known to those skilled in the art such as trifluoroacetic acid in methylene chloride for the tert-butoxycarbonyl protected products.
Compounds of the Formula I wherein R3 is hydroxy, may preferably be prepared by cleavage of a Formula I compound wherein R3 is (C1-C4)alkoxy.
The cleavage reaction may conveniently be carried out by any of the many procedures known for such a transformation. Treatment of the heteroaryl-fused pyrimidine derivative of Formula I with molten pyridine hydrochloride (20-30 eq.) at 150 to 175xc2x0 C. may be employed for O-dealkylations. Alternatively, the reaction may be carried out, for example, by treatment of the heteroaryl-fused pyrimidine derivative with an alkali metal (C1-C4)alkylsulphide such as sodium ethanethiolate or, for example, by treatment with an alkali metal diarylphosphide such as lithium diphenylphosphide. Alternatively the cleavage reaction may conveniently be carried out, for example, by treatment of the heteroaryl-fused pyrimidine derivative with a boron or aluminum trihalide such as boron tribromide. Such reactions are preferably carried out in the presence of a reaction-inert solvent and at a suitable temperature.
For the production of those compounds of Formula I wherein R3 is a (C1-C4)alkylsulphonyl or (C1-C4)alkylsulphonyl group, the oxidation of a Formula I compound wherein R3 is a (C1-C4)alkylthio group Is preferred.
A suitable oxidizing agent is, for example, an agent known in the art for the oxidation of thio to sulphinyl and/or sulphenyl, for example, hydrogen peroxide, a peracid (such as 3-chloroperoxybenzoic or peroxyacetic acid), an alkali metal peroxysulphate (such as potassium peroxymonosulphate), chromium trioxide or gaseous oxygen in the presence of platinum. The oxidation is generally carried out under as mild conditions as possible and with the required stoichiometric amount of oxidizing agent in order to reduce the risk of over oxidation and damage to other functional groups. In general the reaction is carried out in a suitable solvent such as methylene chloride, chloroform, acetone, tetrahydrofuran or ter-butyl methyl ether and at a temperature, for example, xe2x88x9225 to 50xc2x0 C., conveniently at or near ambient temperature, that is in the range of 15 to 35xc2x0 C. When a compound carrying a sulphinyl group is required a milder oxidizing agent may also be used, for example sodium or potassium metaperiodate, conveniently in a polar solvent such as acetic acid or ethanol. It will be appreciated that when a compound of the Formula I containing a (C1-C4)alkylsulphonyl group is required, it may be obtained by oxidation of the corresponding (C1-C4)alkylsulphonyl compound as well as of the corresponding (C1-C4)alkylthio compound.
For the production of those compounds of Formula I wherein R3 is (C2-C4)alkanoylamino or substituted (C2-C4)alkanoylamino, ureido, 3-phenylureido, benzamido, or sulfonamido, the acylation or sulfonylation of a Formula I compound wherein R3 is amino is appropriate.
A suitable acylating agent is, for example, any agent known in the art for the acylation of amino to acylamino, for example an acyl halide (e.g., a (C2-C4)alkanoyl chloride or bromide or a benzoyl chloride or bromide), an alkanoic acid anhydride or mixed anhydride (e.g., (C2-C4)alkanoic acid anhydride such as acetic anhydride or the mixed anhydride formed by the reaction of an alkanoic acid and a (C1-C4)alkoxycarbonyl halide, for example (C1-C4)alkoxycarbonyl chloride, in the presence of a suitable base). For the production of those compounds of Formula I wherein R1 is ureido or 3-phenylureido, a suitable acylating agent is, for example, a cyanate, for example an alkali metal cyanate such as sodium cyanate or, for example, an isocyanate such as phenyl isocyanate. N-sulfonylations may be carried out with suitable sulfonyl halides or sulfonylanhydrides in the presence of a tertiary amine base. In general the acylation or sulfonylation is carried out in a reaction-inert solvent and at a temperature, in the range, for example, xe2x88x9230 to 120xc2x0 C., conveniently at or near ambient temperature.
For the production of those compounds of Formula I wherein R3 is (C1-C4)alkoxy or substituted (C1-C4)alkoxy or R1 is (C1-C4)alkylamino or substituted mono-N- or di-N,N-(C1-C4)alkylamino, the alkylation, preferably in the presence of a suitable base, of a Formula I compound wherein R1 is hydroxy or amino, as appropriate, is preferred.
A suitable alkylating agent is, for example, any agent known in the art for the alkylation of hydroxy to alkoxy or substituted alkoxy, or for the alkylation of amino to alkylamino or substituted alkylamino, for example an alkyl or substituted alkyl halide, for example a (C1-C4)alkyl chloride, bromide or iodide or a substituted (C1-C4)alkyl chloride, bromide or iodide, in the presence of a suitable base in a reaction-inert solvent and at a temperature in the range, for example, 10 to 140xc2x0 C., conveniently at or near ambient temperature.
For the production of those compounds of Formula I wherein R3 is an amino-, oxy- or cyano-substituted (C1-C4)alkyl substituent, the reaction, preferably in the presence of a suitable base, of a Formula I compound wherein R3 is a (C1-C4)alkyl substituent bearing a displaceable group with an appropriate amine, alcohol or cyanide is appropriate.
The reaction is preferably carried out in a reaction-inert solvent or diluent and at a temperature in the range, for example, 10 to 100xc2x0 C., conveniently at or near ambient temperature.
For the production of those compounds of Formula I wherein R3, R5, or R6 is a carboxy substituent or a substituent which includes a carboxy group, the hydrolysis of a Formula I compound wherein R3, R5, R6 is a (C1-C4)alkoxycarbonyl substituent or a substituent which includes a (C1-C4)alkoxycarbonyl group is desirable.
The hydrolysis may conveniently be preformed, for example, under basic conditions such as an alkali metal hydroxide mediated hydrolysis as illustrated in the accompanying Examples.
For the production of those compounds of Formula I wherein R3 is amino, (C1-C4)alkylamino,di-[(C1-C4)alkyl]amino,pyrrolidin-1-yl, piperidino,morpholino,piperazin-1-yl, 4-(C1-C4)alkylpiperazin-1-yl or (C1-C4)alkythio, the reaction, conveniently in the presence of a suitable base, of a Formula I compound wherein R3 is a displaceable group with an appropriate amine or thiol is preferred.
The reaction is preferably carried out in a reaction-inert solvent or diluent and at a temperature in the range, for example, 10 to 180xc2x0 C., conveniently in the range 100 to 150xc2x0 C.
For the production of those compounds of Formula I wherein R3 is 2-oxopyrrolidin-1-yl or 2-oxopiperidin-1-yl, the cyclisation, in the presence of a suitable base, of a Formula I compound wherein R3 is a halo-(C2-C4)alkanoylamino group is convenient.
The reaction is preferably carried out in a reaction-inert solvent or diluent and at a temperature in the range, for example, 10 to 100xc2x0 C., conveniently at or near ambient temperature.
For the production of compounds of Formula I in which R3 is carbamoyl, substituted carbamoyl, alkanoyloxy or substituted alkanoyloxy, the carbamoylation or acylation of a Formula I compound wherein R3 is hydroxy is convenient.
Suitable acylating agents are for example any agent known in the art for acylation of hydroxyaryl moieties to alkanoyloxy aryl. For example, (C2-C4)alkanoyl halides, (C2-C4)alkanoyl anhydrides or mixed anhydrides, and suitable substituted derivatives thereof may be employed typically In the presence of a suitable base. Alternatively, (C2-C4)alkanoic acids or suitably substituted derivatives thereof may be coupled with a Formula I compound wherein R3 is hydroxy with the aid of a condensing agent such as a carbodiimide. For the production of those compounds of Formula I in which R3 is carbamoyl or substituted carbamoyl, suitable carbamoylating agents are for example a cyanate or an alkyl or arylisocyanate, typically in the presence of a suitable base. Alternatively a suitable intermediate such as the chloroformates or imidazolylacarbonyl derivative of a heteroaryl-fused pyrimidine of Formula I in which R3 is hydroxy may be generated, for example by treatment of said derivative with phosgene (or a phosgene equivalent) or carbonyldiimidazole. The resulting intermediate may then be reacted with an appropriate amine or substituted amine to produce the desired carbamoyl derivatives.
For the production of heteroaryl-fused pyrimidine derivatives of Formula I wherein R3 is aminocarbonyl or a substituted aminocarbonyl, the aminolysis of a suitable intermediate derived from a heteroaryl-fused pyrimidine of Formula I in which R3 is carboxy is preferred.
The activation and coupling of a Formula I compound wherein R3 is carboxy may be performed by a variety of methods known to those skilled in the art. Suitable methods include activation of the carboxyl as an acid halide, azide, symmetric or mixed anhydride, or active ester of appropriate reactivity for coupling with the desired amine. Examples of such types of intermediates and their production and use in couplings with amines may be found extensively in the literature; for example M. Bodansky and A. Bodansky, xe2x80x9cThe Practice of Peptide Synthesisxe2x80x9d, Springer,-Verlag, New York, 1984.
The resulting Formula I compounds may be isolated and purified by standard methods, such as solvent removal and recrystallization or chromatography, if desired.
Optionally substituted indole and indolines, useful in the practice of the invention, and methods for their preparation, are described in co-pending U.S. application Ser. No. 08/200,359, incorporated herein by reference. In addition to methods described therein the preparation of various indolines, indoles, oxindoles, and isatins useful as intermediates are further described in xe2x80x9cHeterocyclic Compounds with Indole and Carbazole Systemsxe2x80x9d, W. C. Sumpter and F. M. Miller, in Vol. 8 of xe2x80x9cThe Chemistry of Heterocyclic Compoundsxe2x80x9d Series, Interscience Publishers Inc., N.Y., 1954 and references contained therein.
Substituted anilines, useful in the practice of the invention, and methods for their preparation, are described in co-pending U.S. application Ser. No. 08/413,300 and PCT application no. PCT/IB95/00436, incorporated herein by reference.
Compounds of the formula ZH wherein ZH is QNH2, useful in the practice of the invention, and methods for their preparation, are described in European Patent application serial no. EP 0 496 617 A1 incorporated herein by reference.
Certain compounds of Formula I can exist in solvated as well as unsolvated forms such as, for example, hydrated forms. It is to be understood that the invention encompasses all such solvated as well as unsolvated forms which possess activity against hyperproliferative diseases.
A suitable pharmaceutically-acceptable salt of a heteroaryl-fused pyrimidine derivative of the invention is, for example, an acid-addition salt of a heteroaryl-fused pyrimidine derivative of the invention which is sufficiently basic, for example an acid-addition salt with, for example, an inorganic or organic acid, for example, hydrochloric, hydrobromic, sulphuric, phosphoric, methanesulfonic, benzenesulfonic, trifluoroacetic, citric, lactic or maleic acid. In addition a suitable pharmaceutically-acceptable base-addition salt of a heteroaryl-fused pyrimidine derivative of the invention which is sufficiently acidic is an alkali metal salt, for example a lithium, sodium or potassium salt; an alkaline earth metal salt, for example a calcium or magnesium salt; an ammonium salt; or a salt with an organic base which affords a physiologically-acceptable cation for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine. All such salts are within the scope of this invention and they can be prepared by conventional methods. For example, they can be prepared simply by contacting the acidic and basic entities, usually in a stoichiometric ratio, in either an aqueous, non-aqueous or partially aqueous medium, as appropriate. The salts are recovered either by filtration, by precipitation with a non-solvent, preferably an ethereal or hydrocarbon solvent, followed by filtration, by evaporation of the solvent, or, in the case of aqueous solutions, by lyophilizaton, as appropriate.
Some of the compounds of Formula I have asymmetric carbon atoms. Such diasteromeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods known per se., for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixtures into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., alcohol), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomer. All such isomers, including diastereomers and enantiomers are considered as part of the invention.
The compounds of this invention are potent inhibitors of the erbB family of oncogenic and protooncogenic protein tyrosine kinases such as epidermal growth factor receptor (EGFR), erbB2, HER3, or HER4 and thus are all adapted to therapeutic use as antiproliferative agents (e.g., anticancer) in mammals, particularly humans. In particular, the compounds of this invention are therapeutants or prophylactics for the treatment of a variety of human tumors (renal, liver, kidney, bladder, breast, gastric, ovarian, colorectal, prostate, pancreatic, lung, vulval, thyroid, hepatic carcinomas, sarcomas, glioblastomas, various head and neck tumors), and other hyperplastic conditions such as benign hyperplasia of the skin (e.g., psoriasis) or prostate (e.g., BPH). It is in addition expected that a heteroaryl-fused pyrimidine of the present invention may possess activity against a range of leukemias and lymphoid malignancies.
The compounds of Formula I may also be expected to be useful in the treatment of additional disorders in which aberrant expression ligand/receptor interactions, activation or signalling events related to various protein tyrosine kinases, (e., IGF-receptors) whose activity is inhibited by the agents of Formula I, are involved.
Such disorders may include those of neuronal, glial, astrocytal, hypothalamic, and other glandular, macrophagal, epithelial, stromal, and blastocoelic nature in which aberrant function, expression, activation or signalling of the tyrosine kinases may be involved. In addition, compounds of Formula I may have therapeutic utility in inflammatory, angiogenic and immunologic disorders involving both identified and as yet unidentified tyrosine kinases which are inhibited by compounds of Formula I.
The in vitro activity of these compounds in inhibiting the receptor tyrosine kinase (and thus subsequent proliferative response, e.g., cancer) may be determined by a procedure as detailed below. Activity of compounds of Formula I in vitro can be determined by the amount of inhibition of the phosphorylation of an exogenous substrate (e.g., Lys3xe2x80x94Gastrin or polyGluTyr (4:1) random copolymer (I. Posner et. al., J. Biol. Chem. 267 (29), 20638-47 (1992)) on tyrosine by epidermal growth factor receptor kinase by a test compound relative to a control. Affinity purified, soluble human EGF receptor (96 ng) is obtained according to the procedure in G. N. Gill, W. Weber, Methods in Enzymology 146, 82-88 (1987) from A431 cells (American Type Culture Collection, Rockville, Md.) and preincubated in a microfuge tube with EGF (2 xcexcg/ml) in phosphorylation buffer+ vanadate (PBV: 50 mM HEPES, pH 7.4; 125 mM NaCl; 24 mM MgCl2; 100 xcexcM sodium orthovanadate), in a total volume of 10 xcexcl, for 20-30 minutes at room temperature. The test compound, dissolved in dimethylsulfoxide (DMSO), is diluted in PBV*, and 10 xcexcl is mixed with the EGF receptor /EGF mix, and incubated for 10-30 minutes at 30xc2x0 C. The phosphorylation reaction is initiated by addition of 20 xcexcl 33P-ATP/substrate mix (120 xcexcM Lys3-Gastrin (sequence in single letter code for amino acids, KKKGPWLEEEEEAYGWLDF), 50 mM Hepes pH 7.4, 40 xcexcM ATP, 2 xcexcCi xcex3-[33P]-ATP) to the EGFr/EGF mix and incubated for 20 minutes at room temperature. The reaction is stopped by addition of 10 xcexcl stop solution (0.5 M EDTA, pH 8; 2 mM ATP) and 6 xcexcl 2N HCl. The tubes are centrifuged at 14,000 RPM, 4xc2x0 C., for 10 minutes. 35 xcexcl of supernatant from each tube is pipetted onto a 2.5 cm circle of Whatman P81 paper, bulk washed four times in 5% acetic acid, 1 liter per wash, and then air dried. This results in the binding of substrate to the paper with loss of free ATP on washing. The [33P] incorporated is measured by liquid scintillation counting. Incorporation in the absence of substrate (e.g., lys3-gastrin) is subtracted from all values as a background and percent inhibition is calculated relative to controls without test compound present.
Such assays carried out with a range of doses of test compounds allow the determination of an approximate IC50 value for the in vitro inhibition of EGFR kinase activity. Although the inhibitory properties of the compounds of Formula I vary with structural change as expected, in general, the activity exhibited by these agents determined in the manner described above is in the range of IC50xe2x95x900.0001-30 xcexcM.
Activity of compounds of Formula I in vivo can be determined by the amount of inhibition of tumor growth by a test compound relative to a control. The tumor growth inhibitory effects of various compounds are measured according to the methods of Corbett T. H., et al. xe2x80x9cTumor Induction Relationships in Development of Transplantable Cancers of the Colon in Mice for Chemotherapy Assays, with a Note on Carcinogen Structurexe2x80x9d, Cancer Res., 35, 2434-2439 (1975) and Corbett, T. H., et al., xe2x80x9cA Mouse Colon-tumor Model for Experimental Therapyxe2x80x9d, Cancer Chemother. Rep. (Part 2)xe2x80x9d, 5, 169-186 (1975), with slight modifications. Tumors are induced in the left flank by s.c. injection of 1xc3x97106 log phase cultured tumor cells (human MDA-MB468 breast or human HN5 head and neck carcinoma cells) suspended in 0.10 ml RPMI 1640. After sufficient time has elapsed for the tumors to become palpable. (2-3 mm in diameter) the test animals (athymic mice) are treated with compound (formulated by dissolution in DMSO typically at a concentration of 50 to 100 mg/mL followed by 1:9 dilution into 0.1% Pluronic(copyright) P105 in 0.9% saline) by the intraperitoneal (ip) or oral (po) routes of administration twice daily (i.e., every 12 hours) for 5 to 20 consecutive days. In order to determine an anti-tumor effect, the tumor is measured in millimeters with Vernier calipers across two diameters and the tumor size (mg) is calculated using the formula: Tumor weight=(lengthxc3x97[width]2)/2, according to the methods of Geran, R. I., et al. xe2x80x9cProtocols for Screening Chemical Agents and Natural Products Against Animal Tumors and Other Biological Systemsxe2x80x9d, Third Edition, Cancer Chemother. Results are expressed as percent inhibition, according to the formula: Inhibition (%)=(TuWcontrolxe2x88x92TuWtest)TuWcontrolxc3x97100%. The flank site of tumor implantation provides reproducible dose/response effects for a variety of chemotherapeutic agents, and the method of measurement (tumor diameter) is a reliable method for assessing tumor growth rates.
Administration of the compounds of this invention can be via any method which enables delivery of the compounds to the site of action (e.g., cancer cells). These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical administration, etc.
The amount of heteroaryl-fused pyrimidine derivative administered will, of course, be dependent on the subject being treated, on the severity of the affliction, on the manner of administration and on the judgement of the prescribing physician. However an effective dosage is in the range of approximately 0.1-100 mg/kg, preferably 1 to 35 mg/kg in single or divided doses. For an average 70 kg human, this would amount to 0.05 to 7 g/day, preferably 0.2 to 2.5 g/day.
The composition may, for example, be in a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulations, solution, suspension, for parenteral injection as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository. The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosages. The pharmaceutical compositions will include a conventional pharmaceutical carrier or excipient and a compound according to the invention as an active ingredient. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.
Pharmaceutical compositions according to the invention may contain 0.1%-95% of the compound, preferably 1%-70%. In any event, the composition or formulation to be administered will contain a quantity of a compound according to the invention in an amount effective to alleviate or reduce the signs in the subject being treated, i.e., proliferative diseases, over the course of the treatment.
Exemplary parenteral administration forms include solutions or suspensions of a compound according to the invention Formula I in sterile aqueous solutions, for example aqueous propylene glycol or dextrose solutions are employed. Such dosage forms can be suitably buffered, if desired.
Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. These pharmaceutical compositions can, if desired, contain additional ingredients such as flavorings, binders, excipients and the like. Thus for oral administration, tablets containing various excipients, such as citric acid can be employed, together with various disintegrants such as starch, alginic acid and certain complex silicates and with binding agents such as sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often useful for tableting purposes. Solid compositions of a similar type may also be employed in soft and hard filled gelatin capsules. Preferred materials therefore include lactose or milk sugar and high molecular weight polyethylene glycols. When the aqueous suspensions or elixirs are desired for oral administration the essential active ingredient therein may be combined with various sweetening or flavoring agents, coloring matters or dyes and, if desired, emulsifying agents or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.
Methods of preparing various pharmaceutical compositions with a certain amount of active ingredient are known, or will be apparent, to those skilled in this art. For examples, see Remington""s Pharmaceutical Sciences., Mack Publishing Company, Easter, Pa., 15th Edition (1975).
The anticancer treatment described above may be applied as a sole therapy or may involve, in addition to the heteroaryl-fused pyrimidine derivative of the invention, one or more other antitumor substances. Such conjoint treatment may be achieved by way of the simultaneous, sequential, cyclic or separate dosing of the individual components of the treatment.